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Background: Free-living amoebae (FLA) are protozoa that are widely distributed in the environment mainly in water and soil related habitats. Thermophilic amoebae are among the most common FLA in water bodies, being Vermamoeba vermiformis one of the most common species reported worldwide from these sources. Interestingly, V. vermiformis has often been reported to survive at high temperatures and osmotic pressure worldwide. Materials and Methods: In this study, snow samples were collected from Mount Teide, Tenerife, Canary Islands during the winter season of 2014. The samples were culture on non nutrient agar plates and checked daily for the presence of FLA. After a week, V. vermiformis amoebae were observed in the plates incubated at room temperature and 37°C. Results: Molecular characterization was carried out by amplifying the 18S rDNA gene and DNA sequencing, confirmed that the isolated strain belonged to Vermamoeba vermiformis species. Conclusion: To the best of our knowledge, this is the first report of Vermamoeba vermiformis isolation from such an inusual habitat (low temperatures and high altitude) and the first report of these species in the Canary islands. [ABSTRACT FROM AUTHOR]

The present study describes two cases of Acanthamoeba infections (keratitis and ascites/peritonitis) in small breed domestic dogs in Tenerife, Canary Islands, Spain. In both cases, amoebic trophozoites were observed under the inverted microscope and isolated from the infected tissues and/or fluids, without detecting the presence of other viral, fungal or bacterial pathogens. Amoebae were isolated using 2 % non-nutrient agar plates and axenified for further biochemical and molecular analyses. Osmotolerance and thermotolerance assays revealed that both isolates were able to grow up to 37 °C and 1 M of mannitol and were thus considered as potentially pathogenic. Moreover, the strains were classified as highly cytotoxic as they cause more than 75 % of toxicity when incubated with two eukaryotic cell lines. In order to classify the strains at the molecular level, the diagnostic fragment 3 (DF3) region of the 18S rDNA of Acanthamoeba was amplified and sequenced, revealing that both isolates belonged to genotype T4. In both cases, owners of the animals did not allow any further studies or follow-up and therefore the current status of these animals is unknown. Furthermore, the isolation of these pathogenic amoebae should raise awareness with the veterinary community locally and worldwide. [ABSTRACT FROM AUTHOR]

A 22-month-old male Spanish water dog was hospitalized after its physical examination revealed fever and movement difficulty. After 24 h, the dog was found to have a high fever (39.5 °C) and was treated empirically with doxycycline/ciprofloxacin. At 48 h, after submission the fever rose to 41 °C and the animal presented with a stiff neck and dehydration. Peripheral blood and cerebrospinal fluid (CSF) were sampled and trophozoites with an Acanthamoeba -like morphology were observed in the CSF. PCR specific for Acanthamoeba , Naegleria fowleri and Balamuthia mandrillaris were performed and the CSF sample found positive for Acanthamoeba . Lungs, kidney, liver and spleen samples were collected post mortem. All collected organ samples were positive for Acanthamoeba by PCR, thus confirming a multisystemic infection. Water samples taken at a suspected site of infection yielded an almost identical PCR fragment to those of the clinical samples, indicating that this was probably where the infection originated. This is the first report of a fatal case of Acanthamoeba disseminated infection in a dog in Spain. [ABSTRACT FROM AUTHOR]